Question

1.How would you identify the mutation in PKD1 in the patient samples?

2.How could you introduce the disease mutation into the PKD1 gene

Answer 1

1) Identification oo mutation in PKD1 in the patient samples can be done using mutational analyses of PKD genes in number of patient samples using direct PCR-sequencing and multiplex ligation-dependent probe amplification. Apart from Sanger sequencing, we can also perform next-generation sequencing by pooling long-range PCR amplicons and multiplexing bar-coded libraries.

2) We could introduce the disease mutation into the PKD1 gene by using the primers with mutation to be incorporated and get the mRNA transcript with mutation incorporated. This would be later analyzed by next-generation sequencing analysis whether the mutations are incoporated or not.